LOCUS       pKG01779-pIVT-iC        3369 bp    DNA     circular SYN 10-JAN-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3369)
  AUTHORS   ISU Biology IT
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3369
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     1..39
                     /label=Amp-F-3-UTR
                     /note="color: black; sequence: 
                     gttttgctatgctgtattttacgcggaacccctatttgt; added: 2023-02-09"
     promoter        22..126
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(59..80)
                     /label=oSEQ095 oKL157
                     /note="color: black; sequence: ctcatgagcggatacatatttg; 
                     added: 2023-02-24"
     CDS             join(127..195,196..987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 127 .. 195 / #ccffcc / signal sequence
                        2: 196 .. 987 / #ccffcc
                     Cleavage site after base 195"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGYRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     127..151
                     /label=oSEQ083 Amp-fwd
                     /note="color: black; sequence: atgagtattcaacatttccgtgtcg; 
                     added: 2023-02-24"
     primer_bind     complement(345..364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     primer_bind     complement(345..364)
                     /label=oSEQ066 Amp_rev
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC; added:
                     2023-02-24"
     primer_bind     825..863
                     /label=Amp2-ori_fwd
                     /note="color: black; sequence: 
                     tggagccggtgagcgtgggtaTCGCGGTATCATTGCAGC; added: 2021-01-22"
     primer_bind     complement(827..844)
                     /label=AmpR1_rev
                     /note="color: black; sequence: ACCCACGCTCACCGGCTC; added: 
                     2021-01-22"
     rep_origin      1158..1746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     1647..1666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     1647..1666
                     /label=oSEQ067 pBR322-ori_fwd
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT; added:
                     2023-02-24"
     primer_bind     complement(1729..1791)
                     /label=ori-R-5-UTR
                     /note="color: black; sequence: 
                     gccaaagtgatgggccagcacacagaccagcacgttgcccaggagTTTCCataggctcc
                     gccc; added: 2023-02-09"
     5'UTR           1747..1808
                     /label=beta-globin 5' UTR
                     /note="color: #a6acb3"
     primer_bind     1747..1764
                     /label=eGFP-IVT-F
                     /note="color: black; sequence: 
                     AGCTATAATACGACTCACTATAGGGctcctgggcaacgtgctg; added: 
                     2022-10-28"
     primer_bind     complement(1792..1808)
                     /label=pHA-b-globin-IVT-R
                     /note="color: black; sequence: gaccggtagcgtgcttt; added: 
                     2023-06-06"
     primer_bind     1793..1825
                     /label=recombinase-IVT-F
                     /note="color: black; sequence: 
                     aagcacgctaccggtcACGCGTGCCGCCAccat; added: 2023-06-06"
     misc_feature    1815..1823
                     /label=Kozak
                     /note="color: #00ffff"
     CDS             1824..2879
                     /codon_start=1
                     /label=NLS-iCre
                     /note="color: #ff3985"
                     /translation="MVPKKKRKVSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAF
                     SEHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLN
                     MLHRRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRC
                     QDIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALS
                     LGVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIY
                     GAKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSET
                     GAMVRLLEDGD"
     primer_bind     complement(2883..2917)
                     /label=recombinase-IVT-R
                     /note="color: black; sequence: 
                     ttcgaagcttgagctcgagACCTGAGGAGTGCGGC; added: 2023-06-06"
     primer_bind     2899..2922
                     /label=pHA-b-globin-IVT-F
                     /note="color: black; sequence: ctcgagctcaagcttcgaattcac; 
                     added: 2023-06-06"
     primer_bind     2899..2918
                     /label=3-UTR-F
                     /note="color: black; sequence: ctcgagctcaagcttcgaat; added:
                     2023-02-09"
     3'UTR           join(2996..3369,1..21)
                     /label=beta-globin-3'UTR
                     /note="human beta-globin polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3094..3129)
                     /label=IVT-mRNA-R-from-paper
                     /note="color: black; sequence: 
                     ttttttttttttttttttttttttttttttttttttttttttttttttttttttttttt
                     tttttttttttttttttttttttttttttttttttttttttttttttttttttttttgc
                     aatgaaaataaatgttttttattaggcagaat; added: 2022-10-28"
     primer_bind     complement(join(3363..3369,1..26))
                     /label=3-UTR-R
                     /note="color: black; sequence: 
                     ccgcgtaaaatacagcatagcaaaactttaacc; added: 2023-02-09"
ORIGIN
        1 gttttgctat gctgtatttt acgcggaacc cctatttgtt tatttttcta aatacattca
       61 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      361 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      841 gggtatcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
      961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1741 tggaaactcc tgggcaacgt gctggtctgt gtgctggccc atcactttgg caaagcacgc
     1801 taccggtcac gcgtgccgcc accatggtgc ccaagaagaa gaggaaagtc tccaacctgc
     1861 tgactgtgca ccaaaacctg cctgccctcc ctgtggatgc cacctctgat gaagtcagga
     1921 agaacctgat ggacatgttc agggacaggc aggccttctc tgaacacacc tggaagatgc
     1981 tcctgtctgt gtgcagatcc tgggctgcct ggtgcaagct gaacaacagg aaatggttcc
     2041 ctgctgaacc tgaggatgtg agggactacc tcctgtacct gcaagccaga ggcctggctg
     2101 tgaagaccat ccaacagcac ctgggccagc tcaacatgct gcacaggaga tctggcctgc
     2161 ctcgcccttc tgactccaat gctgtgtccc tggtgatgag gagaatcaga aaggagaatg
     2221 tggatgctgg ggagagagcc aagcaggccc tggcctttga acgcactgac tttgaccaag
     2281 tcagatccct gatggagaac tctgacagat gccaggacat caggaacctg gccttcctgg
     2341 gcattgccta caacaccctg ctgcgcattg ccgaaattgc cagaatcaga gtgaaggaca
     2401 tctcccgcac cgatggtggg agaatgctga tccacattgg caggaccaag accctggtgt
     2461 ccacagctgg tgtggagaag gccctgtccc tgggggttac caagctggtg gagagatgga
     2521 tctctgtgtc tggtgtggct gatgacccca acaactacct gttctgccgg gtcagaaaga
     2581 atggtgtggc tgccccttct gccacctccc aactgtccac ccgggccctg gaagggatct
     2641 ttgaggccac ccaccgcctg atctatggtg ccaaggatga ctctgggcag agatacctgg
     2701 cctggtctgg ccactctgcc agagtgggtg ctgccaggga catggccagg gctggtgtgt
     2761 ccatccctga aatcatgcag gctggtggct ggaccaatgt gaacattgtg atgaactaca
     2821 tcagaaacct ggactctgag actggggcca tggtgaggct gctcgaggat ggggactgag
     2881 cggccgcact cctcaggtct cgagctcaag cttcgaattc accccaccag tgcaggctgc
     2941 ctatcagaaa gtggtggctg gtgtggctaa tgccctggcc cacaagtatc actaagctcg
     3001 ctttcttgct gtccaatttc tattaaaggt tcctttgttc cctaagtcca actactaaac
     3061 tgggggatat tatgaagggc cttgagcatc tggattctgc ctaataaaaa acatttattt
     3121 tcattgcaat gatgtattta aattatttct gaatatttta ctaaaaaggg aatgtgggag
     3181 gtcagtgcat ttaaaacata aagaaatgaa gagctagttc aaaccttggg aaaatacact
     3241 atatcttaaa ctccatgaaa gaaggtgagg ctgcaaacag ctaatgcaca ttggcaacag
     3301 cccctgatgc ctatgcctta ttcatccctc agaaaaggat tcaagtagag gcttgatttg
     3361 gaggttaaa
//
